Enabling non-viral DNA delivery using lipid nanoparticles co-loaded with endogenous anti-inflammatory lipids

Patel, M. N.; Tiwari, S.; Wang, Y.; O'Neill, S.; Wu, J.; Omo-Lamai, S.; Espy, C.; Chase, L. S.; Majumdar, A.; Hoffman, E.; Shah, A.; Sarkozy, A.; Katzen, J.; Pardi, N.; Brenner, J. S.

doi:10.1101/2024.06.11.598533

bioRxiv · 10.1101/2024.06.11.598533

Enabling non-viral DNA delivery using lipid nanoparticles co-loaded with endogenous anti-inflammatory lipids

Patel, M. N.·Tiwari, S.·Wang, Y.·O'Neill, S.·Wu, J.·Omo-Lamai, S.·Espy, C.·Chase, L. S.·Majumdar, A.·Hoffman, E.·Shah, A.·Sarkozy, A.·Katzen, J.·Pardi, N.·Brenner, J. S.

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Abstract

Lipid nanoparticles (LNPs) have transformed genetic medicine, recently shown by their use in COVID-19 mRNA vaccines. While loading LNPs with mRNA has many uses, loading DNA would provide additional advantages such as long-term expression and availability of promoter sequences. However, here we show that plasmid DNA (pDNA) delivery via LNPs (pDNA-LNPs) induces acute inflammation in naive mice which we find is primarily driven by the cGAS-STING pathway. Inspired by DNA viruses that inhibit this pathway for replication, we co-loaded endogenous lipids that inhibit STING into pDNA-LNPs. Specifically, loading nitro-oleic acid (NOA) into pDNA-LNPs (NOA-pDNA-LNPs) ameliorates serious inflammatory responses in vivo enabling prolonged transgene expression (at least 1 month). Additionally, we demonstrate the ability to iteratively optimize NOA-pDNA-LNPs expression by performing a small LNP formulation screen, driving up expression 50-fold in vitro. Thus, NOA-pDNA-LNPs, and pDNA-LNPs co-loaded with other bioactive molecules, will provide a major new tool in the genetic medicine toolbox, leveraging the power of DNAs long-term and promoter-controlled expression.

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Patel, M. N., Tiwari, S., Wang, Y., O'Neill, S., Wu, J., Omo-Lamai, S., Espy, C., Chase, L. S., Majumdar, A., Hoffman, E., Shah, A., Sarkozy, A., Katzen, J., Pardi, N., Brenner, J. S.. 2024-06-12. Enabling non-viral DNA delivery using lipid nanoparticles co-loaded with endogenous anti-inflammatory lipids. https://doi.org/10.1101/2024.06.11.598533

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